Autophagy And Apoptosis In Thyroid Gland Injury Induced By Sleep Deprivation

Objectives 1.In order to study the effect of sleep deprivation on thyroid gland,a sleep deprivation animal model was established,and the electrochemiluminescence method was used to detect thyroid Thyroid hormone..2.To investigate the role and mechanism of autophagy and apoptosis in thyroid injury by detecting autophagy and apoptosis-related genes and proteins in sleep-deprived rats..MethodsAccording to the standard,30 healthy male adult Wistar rats(weighing 300-350g)provided by the animal experimental center of Tianjin Medical University were selected as the experimental objects.They were placed in a standard laboratory animal feeding room,which required 12 hours of light(normal light from 8 a.m.to 20 p.m.)in a day,followed by 12 hours of dark environment,with indoor temperature of 23 ± 1 ℃ and humidity of 55 ± 5%.Provide rats with sufficient food and water.We randomly divided the 30 Wistar rats into three groups(10 rats in each group): normal control group(CC),wide platform control group(TC)and sleep deprivation group(SD).In this experiment,the "small platform water environment" method was used to carry out sleep deprivation experiment in rats.The sleep deprived rats were deprived of sleep for 6 hours(from 8:00to 14:00 in the morning)after entering the sleep deprived experimental box.In order to minimize the non-specific impact of environmental factors,the animals in CC group were not given any treatment after entering the laboratory.Animals in the TC group were placed in the same water tank with a wide platform(160mm diameter),allowing them to eat,move and sleep freely in similar water environment.The other treatment methods of TC group were the same as those of CC group.The experiment lasted for 21 days.21 days later,according to the ethical requirements,rats were anesthetized intravenously,and then blood was collected from the femoral artery of rats.Serum was collected by centrifugation(3000 × g,10min)of the samples in a high centrifuge,and serum biochemical tests were carried out to determine the thyroid function of the rats;part of the thyroid tissues of the rats were paraffin embedded and sectioned,and the histological changes of the thyroid were observed with a microscope.The expression of autophagy related protein and apoptosis related protein(including lc3 a,lc3b,p62,beclin-i,Bax and Bcl-2)in mRNA level and protein level were detected by RT-PCR and Western blotting,and the results were statistically analyzed.Results1.Compared with CC group and TC group,the rats in sleep deprivation group lost weight and had poor mental status.2.The levels of T3,T4,FT3 and FT4 in serum of rats were detected.Compared with CC group,the levels of TT4 in SD group and TC group were significantly higher.The increased secretion of TT3 and TT4 may indicate that sleep deprivation causes the regulation of thyroid axis..3.Histomorphological changes: the observation of thyroid slices suggested that the anatomical structure of thyroid cells in CC and TC group was normal,while some changes occurred in SD group.In the morphology of thyroid cells,the thyroid cells of CC and TC group showed polygon,regular cell arrangement,big and round nucleus,middle,uniform and abundant cytoplasm without vacuole.However,the thyroid cells in SD group showed obvious edema,balloon like or vacuolation degeneration,accompanied by a small number of thyroid cells necrosis,hyperchromatic cytoplasm,nuclear pyknosis,fragmentation or dissolution.4.The results showed that the ratio of TPO and TG in sleep deprivation group was significantly higher than that in CC group and TC group,and the ratio of TG in sleep deprivation group was similar to that in CC group and TC group,but significantly less than that in the former two groups.Compared with CC and TC group,TG MRNA protein level decreased in sleep deprived group,while TPO MRNA protein level increased in sleep deprived group..5.Lc3b(a)II / I ratio in sleep deprivation group was significantly lower than that in CC group.Compared with CC group and TC group,Beclin I protein level decreased and P62 protein level increased in sleep deprivation group,which suggested that sleep deprivation induced autophagy reduction in thyroid cells..6.The expression of Bax and bcl-2 protein changed in different degrees.Compared with CC and TC group,the level of Bax protein in sleep deprivation group increased,while the level of Bcl-2 protein in sleep deprivation group decreased significantly.The expression of Bax / Bcl-2 protein showed that sleep deprivation increased the apoptosis of thyroid.Conclusion1.Sleep deprivation can reduce the weight of thyroid gland and lead to morphological changes.In the sleep deprived group,thyroid cells were damaged and the regulation of thyroid function axis was changed..2.During sleep deprivation,TPO increased while TG decreased.The increase of TPO and the decrease of TG reflect the injury and self-regulation of thyroid.3.The changes of autophagy related proteins suggest that sleep deprivation can induce the decrease of autophagy and the increase of thyroid apoptosis..4.Autophagy and apoptosis play an important role in sleep deprivation.